Frontiers in pediatric testicular torsion: An integrated review of prevailing trends and management outcomes.

Testicular torsion remains the most frequent cause of testicular ischemia, especially in adolescents and young adults. Timely diagnosis and intervention are keys to saving the affected testicle. This review presents current trends in the diagnosis and treatment of torsion, potential pitfalls and consequent outcomes. Additionally, other salient issues surrounding testicular torsion are also discussed, including: pathogenesis of injury, legal ramifications, fertility outcomes, novel management techniques, and recent advances in diagnostic technology.

GPRC5A is a potential oncogene in pancreatic ductal adenocarcinoma cells that is upregulated by gemcitabine with help from HuR.

GPRC5A is an orphan G-protein coupled receptor with an intriguing dual behavior, acting as an oncogene in some cancers and as a tumor suppressor in other cancers. In the pancreatic cancer context, very little is known about GPRC5A. By analyzing messenger RNA (mRNA) expression data from 675 human cancer cell lines and 10 609 samples from The Cancer Genome Atlas (TCGA) we found that GPRC5A's abundance in pancreatic cancer is highest (cell lines) or second highest (TCGA) among all tissues and cancer types. Further analyses of an independent set of 252 pancreatic normal and cancer samples showed GPRC5A mRNA to be more than twofold upregulated in primary tumor samples compared with normal pancreas (P-value<10(-5)), and even further upregulated in pancreatic cancer metastases to various organs (P-value=0.0021). Immunostaining of 208 cores (103 samples) of a tissue microarray showed generally low expression of GPRC5A protein in normal pancreatic ductal cells; on the other hand, in primary and metastatic samples, GPRC5A protein levels were dramatically increased in pancreatic ductal cells. In vitro studies of multiple pancreatic cancer cell lines showed that an increase in GPRC5A protein levels promoted pancreatic cancer cell growth and migration. Unexpectedly, when we treated pancreatic cancer cell lines with gemcitabine (2',2'-difluorodeoxycytidine), we observed an increase in GPRC5A protein abundance. On the other hand, when we knocked down GPRC5A we sensitized pancreatic cancer cells to gemcitabine. Through further experimentation we showed that the monotonic increase in GPRC5A protein levels that we observe for the first 18 h following gemcitabine treatment results from interactions between GPRC5A's mRNA and the RNA-binding protein HuR, which is an established key mediator of gemcitabine's efficacy in cancer cells. As we discovered, the interaction between GPRC5A and HuR is mediated by at least one HuR-binding site in GPRC5A's mRNA. Our findings indicate that GPRC5A is part of a complex molecular axis that involves gemcitabine and HuR, and, possibly, other genes. Further work is warranted before it can be established unequivocally that GPRC5A is an oncogene in the pancreatic cancer context.

The mRNA-binding protein HuR promotes hypoxia-induced chemoresistance through posttranscriptional regulation of the proto-oncogene PIM1 in pancreatic cancer cells.

Previously, it has been shown that pancreatic ductal adenocarcinoma (PDA) tumors exhibit high levels of hypoxia, characterized by low oxygen pressure (pO2) and decreased O2 intracellular perfusion. Chronic hypoxia is strongly associated with resistance to cytotoxic chemotherapy and chemoradiation in an understudied phenomenon known as hypoxia-induced chemoresistance. The hypoxia-inducible, pro-oncogenic, serine-threonine kinase PIM1 (Proviral Integration site for Moloney murine leukemia virus 1) has emerged as a key regulator of hypoxia-induced chemoresistance in PDA and other cancers. Although its role in therapeutic resistance has been described previously, the molecular mechanism behind PIM1 overexpression in PDA is unknown. Here, we demonstrate that cis-acting AU-rich elements (ARE) present within a 38-base pair region of the PIM1 mRNA 3'-untranslated region mediate a regulatory interaction with the mRNA stability factor HuR (Hu antigen R) in the context of tumor hypoxia. Predominantly expressed in the nucleus in PDA cells, HuR translocates to the cytoplasm in response to hypoxic stress and stabilizes the PIM1 mRNA transcript, resulting in PIM1 protein overexpression. A reverse-phase protein array revealed that HuR-mediated regulation of PIM1 protects cells from hypoxic stress through phosphorylation and inactivation of the apoptotic effector BAD and activation of MEK1/2. Importantly, pharmacological inhibition of HuR by MS-444 inhibits HuR homodimerization and its cytoplasmic translocation, abrogates hypoxia-induced PIM1 overexpression and markedly enhances PDA cell sensitivity to oxaliplatin and 5-fluorouracil under physiologic low oxygen conditions. Taken together, these results support the notion that HuR has prosurvival properties in PDA cells by enabling them with growth advantages in stressful tumor microenvironment niches. Accordingly, these studies provide evidence that therapeutic disruption of HuR's regulation of PIM1 may be a key strategy in breaking an elusive chemotherapeutic resistance mechanism acquired by PDA cells that reside in hypoxic PDA microenvironments.

Bioactive molecules from sea hares.

Sea hares, belonging to the order Opisthobranchia, subclass Gastropoda, are mollusks that have attracted many researchers who are interested in the chemical defense mechanisms of these soft and "shell-less" snails. Numbers of small molecules of dietary origin have been isolated from sea hares and some have ecologically relevant activities, such as fish deterrent activity or toxicity. Recently, however, greater attention has been paid to biomedically interesting sea hare isolates such as dolastatins, a series of antitumor peptide/macrolides isolated from Dolabella auricularia. Another series of bioactive peptide/macrolides, as represented by aplyronines, have been isolated from sea hares in Japanese waters. Although earlier studies indicated the potent antitumor activity of aplyronines, their clinical development has never been conducted because of the minute amount of compound available from the natural source. Recent synthetic studies, however, have made it possible to prepare these compounds and analogs for a structure-activity relationship study, and started to uncover their unique action mechanism towards their putative targets, microfilaments. Here, recent findings of small antitumor molecules isolated from Japanese sea hares are reviewed. Sea hares are also known to produce cytotoxic and antimicrobial proteins. In contrast to the small molecules of dietary origin, proteins are the genetic products of sea hares and they are likely to have some primary physiological functions in addition to ecological roles in the sea hare. Based on the biochemical properties and phylogenetic analysis of these proteins, we propose that they belong to one family of molecule, the "Aplysianin A family," although their molecular weights are apparently divided into two groups. Interestingly, the active principles in Aplysia species and Dolabella auricularia were shown to be L-amino acid oxidase (LAAO), a flavin enzyme that oxidizes an alpha-amino group of the substrate with molecular oxygen and liberates hydrogen peroxide, with a sequence similar to other known LAAOs, including snake venom. Possible antibacterial activity and cytotoxic activity mechanisms of these proteins are also discussed.

Cloning of the Microcystis aeruginosa M228 lectin (MAL) gene.

We have cloned and characterized the gene encoding Microcystis aeruginosa (strain M228) lectin (MAL). The gene contains 1551 nucleotides and an open reading frame for a protein of 517 amino acids with a predicted molecular weight of 55,159 Da. The carboxy-terminal region of MAL has three tandemly repeated homologous domains composed of 61 amino acids. These regions show similarity to the corresponding regions of the alpha-amylase of Clostridium beijerinckii (23% identity). The mal gene lies adjacent to an ORF that display homology to cytochrome P-450 and polyketide synthase. Southern hybridization showed that the genomic DNA of the strain M228 contained, in addition to MAL gene (mal), at least two other mal like gene.

The D-galactose-binding lectin of the octocoral Sinularia lochmodes: characterization and possible relationship to the symbiotic dinoflagellates.

A D-galactose binding lectin (SLL-2) was isolated from Sinularia lochmodes, an octocoral, by a combination of affinity chromatography on acid-treated agarose and FPLC on Superdex 200. SLL-2 agglutinated rabbit and horse erythrocytes while SLL-1, a minor component, reacted only with rabbit erythrocytes. SLL-2 is a glycoprotein with a molecular mass of 122 kDa and is composed of eight identical subunits (15 kDa). The sequence of the amino terminal region of SLL-2 did not show any apparent homology to the sequences of other animal and plant lectins. D-Galactose, N-acetyl-D-galactosamine, lactose, and melibiose were moderate inhibitors to the agglutination of rabbit erythrocytes. In contrast, horse erythrocytes were much more susceptible to agglutination by SLL-2, which was inhibited by sugars and glycoproteins such as D-galactose, N-acetyl-D-galactosamine, lactose, melibiose, and porcine stomach mucin. SLL-2 showed considerable tolerance to heating and kept its activity after heating at 80 degrees C for 60 min. In immuno-histochemical studies using an anti-SLL-2 antiserum and protein A gold conjugate, SLL-2 was found to be present in high amounts in the nematocysts. SLL-2 was also detected on the surface of symbiotic dinoflagellate, Symbiodinium sp. cells irrespective whether they were surrounded with or without host cells. These observations suggest the presence of lectin-mediated interaction between symbiotic dinoflagellates and S. lochmodes.

Development of a new inhibitor of glucosylceramide synthase.

Analogs of the potent inhibitor of glucosylceramide (GlcCer) synthase, D-threo-1-phenyl-2-palmitoylamino-3-pyrrolidino-1-propanol (P4), based on substitutions in the palmitoyl group were made by means of a stereo-selective synthetic method in order to elucidate the role of the hydrophobic portion in both the inhibitory action toward the enzyme and the biological effects. While P4 strongly inhibited GlcCer synthase with an IC(50) of 0.5 microM in vitro, it also inhibited cell growth by 50% at the concentration of 7 microM. The shorter N-acyl chain analogs including decanoyl, octanoyl, and hexanoyl groups showed similar IC(50) values for GlcCer synthase (around 2 microM) but the hexanoyl analog exhibited only a slight inhibitory effect on cell growth, showing the dissociation between GlcCer depletion and cell growth. Several compounds which exhibit similar hydrophobicity to the hexanoyl analog of P4 were subsequently designed. We found that D-threo-1-phenyl-2-benzyloxycarbonylamino-3-pyrrolidino-1-pr opanol (PBPP) was a most potent inhibitor, showing an IC50 of 0.3 microM. In cultured cells, PBPP was able to deplete glycosphingolipids without affecting cell growth or the ceramide level.

Isolation and characterization of a mannan-binding lectin from the freshwater cyanobacterium (blue-green algae) Microcystis viridis.

Microcystis viridis NIES-102 strain, a unicellular freshwater bloom-forming cyanobacterium, showed transient hemagglutinating activity in laboratory culture during stationary phase under nonaeration conditions. However, the hemagglutinating activity which was inhibited with yeast mannan could not be observed during culture with aeration. A mannan-binding lectin named MVL was isolated with the assay of the hemagglutinating activity against rabbit erythrocytes from the cyanobacterium by successive hydrophobic and gel filtration chromatography. MVL was composed of a single polypeptide of 13 kDa. The gene (mvl) for MVL was cloned from a genomic DNA of NIES-102 strain as a template, and its sequence was determined. The deduced amino acid sequence showed that MVL consisted of 113 amino acid residues and was composed of two tandemly repeated homologous domains of 54 amino acid residues. MVL showed no sequence homology to any other lectins or proteins.

Chemical modification of beta-glucocerebrosidase inhibitor N-octyl-beta-valienamine: synthesis and biological evaluation of N-alkanoyl and N-alkyl derivatives.

Several N-alkanoyl 4a-d and N-alkyl derivatives 5a-g of the potent beta-glucocerebrosidase inhibitor N-octyl beta-valienamine (3) were synthesized in order to elucidate a role of hydrophobic portion in the inhibitory action. Although the former lacked inhibitory potency, the latter were strong beta-glucocerebrosidase inhibitors (cf. N-decyl-N-octyl-beta-valienamine 5d: Ki 6.6 x 10(-8) M). Furthermore, when being prescribed into mouse-derived B16 melanoma cells, N-butyl-N-octyl-beta-valienamine 5a and 5d were shown to change the amount of GlcCer and GM3, which suggests that they are possibly introduced into cells and influence glycolipids biosynthesis.

Inhibitory effects of catecholamines and maternal stress on aromatase activity in the fetal rat brain.

OBJECTIVE: Aromatization in the the fetal brain is thought to be involved both in sex differentiation during early development and in adult sexual behavior. Although recently the relationship between aromatase and catecholamine has been discussed, the effect of stress on aromatase in the fetal brain has not been clarified. Therefore, in the present study, localization of aromatase and the inhibitory effects of catecholamines and maternal stress on aromatase activity in the fetal rat brain were examined. METHODS: Localization of aromatase cytochrome P-450 using a specific polyclonal antiserum against human placenta aromatase was examined, and the inhibitory effects of dopamine and norepinephrine on aromatase activity in vitro were studied. Further, the influences of intrauterine stress on aromatase activity in the prenatal rat brain were evaluated in vivo. RESULTS: Aromatase-immunoreactive neurons are located principally in the medial amygdaloid nucleus. Aromatase activity in the fetal rat brain was competitively inhibited by dopamine and norepinephrine, with Ki values of 120 microM and 100 microM, respectively. Aromatase activity in the fetal brain was significantly lower in stressed rats given 1.5% salt water (89.2 +/- 17.5 fmol/mg/hr; n = 4) (p < 0.05) than in the control group (123.1 +/- 10.0 fmol/mg/hr; n = 4). CONCLUSION: Aromatase activity in the prenatal rat brain is influenced by catecholamine metabolism during intrauterine stress.

Synthesis and evaluation of morpholino- and pyrrolidinosphingolipids as inhibitors of glucosylceramide synthase.

This paper describes the new synthesis and evaluation of some morpholino- and pyrrolidinosphingolipids and mimics as inhibitors of glucosylceramide synthase. It was found that the pyrrolidino derivatives are generally more active than the morpholino derivatives and the best one was shown to be a nanomolar inhibitor.

Late cyst convolution after gamma knife radiosurgery for cerebral arteriovenous malformations.

Although many series of patients with cerebral arteriovenous malformations (AVMs) treated radiosurgically have been published, there has been little information on cysts appearing several years after irradiation. Herein, we discuss the incidence, mechanisms and predictability of late cyst formation based on our personal experiences, as well as reported patients. The incidence of this complication, though generally considered to be 0.5% or less, may be higher than assumed. Although a breakdown of the blood-brain barrier is likely to play a major role in the formation process, the hematoma cavity itself may have the potential to become a cyst. A radiation-induced lesion appearing several years after irradiation and persisting for several years thereafter may be a warning sign of late cyst formation. Long-term follow-up, particularly using neuroimaging techniques, is necessary even after the 'treatment goal' has been achieved.

Purification and characterization of Microcystis aeruginosa (freshwater cyanobacterium) lectin.

Microcystis aeruginosa, strain M228, a laboratory culture of freshwater cyanobacterium, showed hemagglutinating activity against rabbit, horse and human ABO erthrocytes. Crossed absorption tests revealed the presence of a single type of lectin in the extract of M228 strain cells. The lectin, termed MAL, was purified in combination with the affinity chromatography on acid-treated agarose gel and the gel permeation chromatography in an electrophoretically pure form. MAL was a glycoprotein containing 7.8% neutral sugars and was composed of a single polypeptide having a molecular weight of 57 kDa. Isoelectric point was estimated to be pH 6.4. Hemagglutinating activity of the lectin was inhibited effectively by N-acetyl-D-galactosamine and by glycoproteins. D-galactose and lactose also showed moderate inhibitory activity. The destruction of the hemagglutinating activity by a 2-mercaptoethanol treatment suggests the presence of intra-chain disulfide bond(s) essential for the activity in the molecule. The sequence of the amino-terminal region of MAL was determined as Val-Leu-Ala-Ser-Leu-Val-Ser-Thr-Ser-Gln-Ala-Gly-Ser-Leu-Glu-Leu-Leu- Ala [corrected].

A synthetic ceramide analog (L-PDMP) up-regulates neuronal function.

To address the role of brain gangliosides in synaptic activity, the ceramide analogs, D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP) and its enantiomer, L-PDMP, were used to inhibit and stimulate ganglioside biosynthesis in cultured cortical neurons. Prolonged treatment with both PDMP isomers exhibited opposite effects on functional synapse formation measured by spontaneous synchronized oscillatory activity of intracellular Ca2+ between the neurons: suppression by D-PDMP and facilitation by L-PDMP. Up-regulation of synaptic activity by L-PDMP could be correlated with the slow but robust activation of p42 mitogen-activated protein kinase. Treatment with L-PDMP after transient forebrain ischemia in rats ameliorated the deficit of a well-learned spatial memory by an 8-arm maze task, suggesting a new potential therapeutic approach for neurodegenerative disorders.

Conservative management of acoustic neurinomas: prospective study of long-term changes in tumor volume and auditory function.

Recently reported retrospective analysis on the natural courses of acoustic neurinomas have disclosed that 26-86% of tumors show very slow, or no, growth for many years. To our knowledge, however, there have been no prospective analyses of the natural course of these tumors. We prospectively analyzed tumor growth as well as auditory function changes in 12 patients (13 tumors) managed conservatively. All 12 patients were advised to undergo both pure tone audiometry and magnetic resonance imaging at 3-4 month intervals for the first follow-up year and at 1-2 year intervals thereafter. Then, in the event of either significant tumor growth (> 20% volume increase) or hearing deterioration (> 10 dB), a decision would be made as to whether the patient should undergo either surgery or radiosurgery. A significant increase in tumor volume was confirmed in seven of the 13 tumors during the mean observation period of 564 days (88-1269 days). In another patient, though neither tumor growth nor worsening of auditory acuity was significant, the patient complained of subjective change in the symptoms. Therefore, we decided to treat eight (62%) of the 13 tumors. In contrast, neither tumor growth nor worsening of auditory acuity has as yet occurred in the remaining five tumors (38%) which have been observed for a mean period of 627 days (342-1377 days). Careful follow-up of these five patients is ongoing. Although further long-term follow-up is clearly necessary, conservative treatment appears to be a reasonable alternative to immediate treatment for selected patients with acoustic neurinomas.

Development of a 3D CT-scanner using a cone beam and video-fluoroscopic system.

We describe the design and implementation of a system that acquires three-dimensional (3D) data of high-contrast objects such as bone, lung, and blood vessels (enhanced by contrast agent). This 3D computed tomography (CT) system is based on a cone beam and video-fluoroscopic system and yields data that is amenable to 3D image processing. An X-ray tube and a large area two-dimensional detector were mounted on a single frame and rotated around objects in 12 seconds. The large area detector consisted of a fluorescent plate and a charge coupled device (CCD) video camera. While the X-ray tube was rotated around the object, a pulsed X-ray was generated (30 pulses per second) and 360 projected images were collected in a 12-second scan. A 256 x 256 x 256 matrix image was reconstructed using a high-speed parallel processor. Reconstruction required approximately 6 minutes. Two volunteers underwent scans of the head or chest. High-contrast objects such as bronchial, vascular, and mediastinal structures in the thorax, or bones and air cavities in the head were delineated in a "real" 3D format. Our 3D CT-scanner appears to produce data useful for clinical imaging and 3D image processing.

Radiation-related adverse effects observed on neuro-imaging several years after radiosurgery for cerebral arteriovenous malformations.

BACKGROUND: To our knowledge, there are no reported arteriovenous malformation (AVM) series in which detailed long-term follow-up results after radiosurgery were described based on the whole patient group. METHOD: We performed a detailed long-term follow-up study of 53 patients with cerebral AVMs treated with gamma knife (GK) radiosurgery, with emphasis on radiation-related adverse effects detected on neuro-imaging after a long post-irradiation latency period (3-10 years). The post-GK follow-up period was 40-232 months excluding two mortalities, the mean being 112 and the median being 111 months. RESULTS: Three patients (5.6%) have, as yet, refused all neuro-imaging follow-up studies. Complete nidus obliteration was confirmed angiographically in 32 patients (60.4%) between 1 and 5 post-GK years. In the other 18 patients (34%), despite significant nidus shrinkage being angiographically demonstrated, complete obliteration was not achieved during a 2-7 year follow-up period. There were two mortalities, one AVM-related (massive re-bleeding during the latency period) and the other angiography-related. There were five radiation-related morbidities (9.4%), three of which-hemi-Parkinson syndrome, hemiparesis, and visual field disturbances attributable to delayed cyst formation-manifested at 5.5, 7 and 7 post-GK years, respectively. We also experienced five patients (9.4%) in whom, despite remaining asymptomatic to date, radiation-related adverse effects were seen on neuro-imaging: middle cerebral artery stenosis at 3 post-GK years in one patient; dural arteriovenous fistula at 7 post GK-years in one; delayed cyst formation in two, at 5 and 10 post-GK years; and a small cavitation at 9 post-GK years. CONCLUSION: Long-term follow-up, particularly with neuro-imaging modalities, is essential even after the "treatment goal" has been attained.

A diffuse white matter ischemic lesion appearing 7 years after stereotactic radiosurgery for cerebral arteriovenous malformations: case report.

OBJECTIVE AND IMPORTANCE: Little information is available about radiation-induced complications occurring more than 5 years after radiosurgical treatment for arteriovenous malformations. CLINICAL PRESENTATION: We present a patient with arteriovenous malformations who experienced hemimotor weakness caused by a diffuse white matter necrotic lesion developing 7 years after gamma knife radiosurgery. The original nidus had been too large (24.1 cm3) to be totally covered and irradiated with a peripheral dose of 20 to 25 Gy. Therefore, the lower half of the nidus, which was adjacent to the major feeding artery, had been partially covered with a 30% isodose volume using two target points with an 18-mm collimator. A central dose of 70 Gy was used to obtain 21 Gy at the periphery. Complete nidus obliteration was angiographically confirmed 38 months after radiosurgery. After a 6-year uncomplicated period, this patient experienced a convulsive seizure and then mild right hemiparesis. INTERVENTION: Computed tomography demonstrated a diffuse hypodense area in the left white matter, which had not been revealed by the previous examination. With steroid treatment, this patient achieved clinical improvement, although there was no significant improvement in the computed tomography-demonstrated white matter lesion. CONCLUSION: Although the evaluation of this patient may not be sufficient and further examinations may be necessary, we tentatively conclude that the computed tomography-demonstrated hypodense lesion in this patient is a radiation-related necrotic lesion. Long-term follow-up is crucial, even after the "treatment goal" has been achieved.